Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish

Dong, Jiexian; Li, Zhenfeng; Wang, Yu; Jin, Maojun; Shen, Yudong; Xu, Zhenlin; Hassıbelnaby, Abdelaty Mostafa Abdelaty; Gee, Shirley J.; Hammock, Bruce D.; Sun, Yuanming; Wang, Hong

doi:10.1016/j.foodchem.2020.127780

Makale Tam Makale Uluslararası 2021

Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish

Jiexian Dong, Zhenfeng Li, Yu Wang, Maojun Jin, Yudong Shen, Zhenlin Xu, Abdelaty Mostafa Abdelaty Hassıbelnaby, Shirley J. Gee, Bruce D. Hammock, Yuanming Sun, Hong Wang

FOOD CHEMISTRY Cilt 337 · 01.02.2021

DOI ile aç

Özet

© 2020 Elsevier LtdTo determine malachite green (MG) and its major metabolite, leucomalachite green (LMG) residual levels in tilapia fish, chemiluminescent enzyme immunoassay (CLEIA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. At first, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against LMG, and then thoroughly by database-assisted sequence analysis. Finally, the productive VH and VL were assembled to an intact scFv sequence and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step CLEIA against LMG was developed with a half-maximal inhibitory concentration (IC50) and limit of detection (LOD) of 1.3 and 0.04 ng/mL, respectively. The validation results of this novel competitive CLEIA was in line with those obtained by classical HPLC method for determination of total MG in spiked and field incurred samples.

Anahtar kelimeler

malachite green residue determination Database-assisted sequence analysis ScFv-AP based immunoassay Tilapia fish

Atıf için künye

Dong, J., Li, Z., Wang, Y., Jin, M., Shen, Y., Xu, Z., Hassıbelnaby, A. M. A., Gee, S. J., Hammock, B. D., Sun, Y., & Wang, H. (2021). Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish. FOOD CHEMISTRY, 337. https://doi.org/10.1016/j.foodchem.2020.127780

BibTeX

@article{dong2021-209550,
  author = {Dong, Jiexian and Li, Zhenfeng and Wang, Yu and Jin, Maojun and Shen, Yudong and Xu, Zhenlin and Hassıbelnaby, Abdelaty Mostafa Abdelaty and Gee, Shirley J. and Hammock, Bruce D. and Sun, Yuanming and Wang, Hong},
  title = {Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish},
  journal = {FOOD CHEMISTRY},
  year = {2021},
  volume = {337},
  issn = {0308-8146},
  doi = {10.1016/j.foodchem.2020.127780},
  url = {https://akademik.atauni.edu.tr/yayinlar/209550-generation-of-functional-single-chain-fragment-variable-from-hybridoma-and}
}